Structural highlights
Publication Abstract from PubMed
Fab is a promising format for antibody drug. Therefore, efforts have been made to improve its thermal stability for therapeutic and commercial use. So far, we have attempted to introduce a disulfide bond into the Fab fragment to improve its thermal stability and demonstrated that it is possible to do this without sacrificing its biochemical function. In this study, to develop a novel stabilization strategy for Fab, we attempted to introduce a disulfide bond between the variable and constant domains and prepared three variants of Fab; H:G10C + H:P210C, L:P40C + L:E165C, and H:G10C + H:P210C + L:P40C + L:E165C. Differential scanning calorimetry measurements showed that each of these variants had improved thermal stability. In addition, the variants with two disulfide bonds demonstrated a 6.5 degrees C increase in their denaturation temperatures compared to wild-type Fab. The introduction of disulfide bonds was confirmed by X-ray crystallography, and the variants retained their antigen-binding activity. The variants were also found to be less aggregative than the wild type. Our results demonstrate that the introduction of a disulfide bond between the variable and constant domains significantly improves the thermal stability of Fab.
Stabilization of adalimumab Fab through the introduction of disulfide bonds between the variable and constant domains.,Yoshikawa M, Senda M, Nakamura H, Oda-Ueda N, Ueda T, Senda T, Ohkuri T Biochem Biophys Res Commun. 2024 Mar 12;700:149592. doi: , 10.1016/j.bbrc.2024.149592. Epub 2024 Jan 28. PMID:38295648[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Yoshikawa M, Senda M, Nakamura H, Oda-Ueda N, Ueda T, Senda T, Ohkuri T. Stabilization of adalimumab Fab through the introduction of disulfide bonds between the variable and constant domains. Biochem Biophys Res Commun. 2024 Mar 12;700:149592. PMID:38295648 doi:10.1016/j.bbrc.2024.149592
